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首页> 外文期刊>Applied and Environmental Microbiology >Purification and Characterization of a Novel Alcohol Dehydrogenase from Leifsonia sp. Strain S749: a Promising Biocatalyst for an Asymmetric Hydrogen Transfer Bioreduction
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Purification and Characterization of a Novel Alcohol Dehydrogenase from Leifsonia sp. Strain S749: a Promising Biocatalyst for an Asymmetric Hydrogen Transfer Bioreduction

机译：一种新型的Leifsonia sp。醇脱氢酶的纯化和鉴定。菌株S749：不对称氢转移生物还原的有前途的生物催化剂。

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摘要

To find microorganisms that could reduce phenyl trifluoromethyl ketone (PTK) to (S)-1-phenyltrifluoroethanol [(S)-PTE], styrene-assimilating bacteria (ca. 900 strains) isolated from soil samples were screened. We found that Leifsonia sp. strain S749 was the most suitable strain for the conversion of PTK to (S)-PTE in the presence of 2-propanol as a hydrogen donor. The enzyme corresponding to the reaction was purified homogeneity, characterized and designated Leifsonia alcohol dehydrogenase (LSADH). The purified enzyme had a molecular weight of 110,000 and was composed of four identical subunits (molecular weight, 26,000). LSADH required NADH as a cofactor, showed little activity with NADPH, and reduced a wide variety of aldehydes and ketones. LSADH catalyzed the enantioselective reduction of some ketones with high enantiomeric excesses (e.e.): PTK to (S)-PTE (>99% e.e.), acetophenone to (R)-1-phenylethanol (99% e.e.), and 2-heptanone to (R)-2-heptanol (>99% e.e.) in the presence of 2-propanol without an additional NADH regeneration system. Therefore, it would be a useful biocatalyst.
机译：为了找到可将苯基三氟甲基酮（PTK）还原为（S）-1-苯基三氟乙醇[（S）-PTE]的微生物，筛选了从土壤样品中分离的苯乙烯同化细菌（约900株）。我们发现Leifsonia sp。在2-丙醇作为氢供体的存在下，菌株S749是最适合PTK转化为（S）-PTE的菌株。纯化对应于该反应的酶同质性，表征并命名为Leifsonia醇脱氢酶（LSADH）。纯化的酶的分子量为110,000，由四个相同的亚基组成（分子量为26,000）。 LSADH需要NADH作为辅助因子，与NADPH几乎没有活性，并且可以还原多种醛和酮。 LSADH催化一些对映体过量对映体（ee）的酮的对映选择性还原：将PTK还原为（S）-PTE（> 99 ％ee），将苯乙酮还原为（R）-1-苯基乙醇（99 ％ee），以及2-在2-丙醇存在下，将庚酮转化为（R）-2-庚醇（> 99％ee），而无需额外的NADH再生系统。因此，它将是有用的生物催化剂。

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